Noriyoshi Manabe; Takuya Sakurai; Fumiko Kihara-Negishi; Yosuke Nakazawa; Toshiyuki  Yamada; Atsushi Iwama; Tsuneyuki Oikawa

doi:10.14715/cmb/2025.71.3.6

Issue

Vol. 71 No. 3: Issue 3

Issue Published : April 15, 2025

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Antagonistic effects of PU.1 on Gfi-1B-induced erythroid colony formation in human cord blood cells

Antagonistic effects of PU.1 on Gfi-1B

https://doi.org/10.14715/cmb/2025.71.3.6

Noriyoshi Manabe

Division of Structural Glycobiology Institute of Molecular Biomembrane and Glycobiology, Tohoku Medical and Pharmaceutical University, Sendai, Japan

Takuya Sakurai

Department of Microbiology and Molecular Cell Biology, Nihon Pharmaceutical University, Saitama, Japan

Fumiko Kihara-Negishi

Department of Life and Health Sciences, School of Pharma-Sciences, Teikyo University, Tokyo, Japan

Yosuke Nakazawa

Division of Hygienic Chemistry, Faculty of Pharmacy, Keio University, Tokyo, Japan

Toshiyuki Yamada

Department of Microbiology and Molecular Cell Biology, Nihon Pharmaceutical University, Saitama, Japan

Atsushi Iwama

Division of Stem Cell and Molecular Medicine, Center for Stem Cell Biology and Regenerative Medicine, The Institute of Medical Science, The University of Tokyo, Tokyo, Japan

Tsuneyuki Oikawa

Medical Genetics Section, Department of Communication Disorders, Psychological Science, Health Sciences University of Hokkaido, Sapporo, Japan

Corresponding Author(s) : Takuya Sakurai

t-sakurai@nichiyaku.ac.jp

Cellular and Molecular Biology, Vol. 71 No. 3: Issue 3
Article Published : April 15, 2025

Abstract

Gfi-1B is a hematopoietic transcription factor essential for growth and differentiation of the erythroid/megakaryocytic lineages, and PU.1 is a master regulator for myeloid development. Herein, we demonstrate that PU.1 interacted with Gfi-1B in vivo by immunoprecipitation assay. GST pull-down assays showed that the binding sites were located in the Ets domain of PU.1 and the zinc finger domain of Gfi-1B. Luciferase reporter assays revealed that PU.1 and Gfi-1B antagonized each other’s transcriptional activity in a dose-dependent manner. The transduction of Gfi-1B alone in human cord blood progenitor cells strongly enhanced erythroid colony formation. However, the transduction of PU.1 along with Gfi-1B in the progenitors significantly inhibited erythroid colony formation. Co-expression of Gfi-1B with a mutant PU.1, which bound to Gfi-1B but not to GATA1, another erythroid master regulator, also inhibited Gfi-1B-induced erythroid colony formation. Our results suggest that the function of Gfi-1B in the growth and differentiation of erythroid cells is antagonized by the expression of PU.1.

Keywords

Transcription Factor Protein-protein interaction Antagonistic effects Hematopoietic differentiation Human cord blood

Manabe, N., Sakurai, T., Kihara-Negishi, F., Nakazawa, Y., Yamada, T., Iwama, A., & Oikawa, T. (2025). Antagonistic effects of PU.1 on Gfi-1B-induced erythroid colony formation in human cord blood cells: Antagonistic effects of PU.1 on Gfi-1B. Cellular and Molecular Biology, 71(3), 48–56. https://doi.org/10.14715/cmb/2025.71.3.6